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Volume 271, Number 23,
Issue of June 7, 1996
pp. 13875-13881
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
Purification and Characterization of Two Isoenzymes of
DL-Glycerol-3-phosphatase from Saccharomyces
cerevisiae
IDENTIFICATION OF THE CORRESPONDING GPP1 AND
GPP2 GENES AND EVIDENCE FOR OSMOTIC REGULATION OF Gpp2p
EXPRESSION BY THE OSMOSENSING MITOGEN-ACTIVATED PROTEIN KINASE SIGNAL
TRANSDUCTION PATHWAY
(Received for publication, January 25, 1996, and in revised form, March 7, 1996)
Joakim
Norbeck
,
Anna-Karin
Påhlman
,
Noreen
Akhtar
,
Anders
Blomberg
and
Lennart
Adler
From the Department of General and Marine Microbiology, Lundberg
Laboratory, Göteborg University, Medicinaregatan 9C,
S-41390 Göteborg, Sweden
The existence of specific
DL-glycerol-3-phosphatase (EC) activity in
extracts of Saccharomyces cerevisiae was confirmed by
examining strains lacking nonspecific acid and alkaline phosphatase
activities. During purification of the glycerol-3-phosphatase, two
isozymes having very similar molecular weights were isolated by gel
filtration and anion exchange chromatography. By microsequencing of
trypsin-generated peptides the corresponding genes were identified as
previously sequenced open reading frames of unknown function. The two
genes, GPP1 (YIL053W) and GPP2 (YER062C) encode
proteins that show 95% amino acid identity and have molecular masses
of 30.4 and 27.8 kDa, respectively. The intracellular concentration of
Gpp2p increases in cells subjected to osmotic stress, while the
production of Gpp1p is unaffected by changes of external osmolarity.
Both isoforms have a high specificity for
DL-glycerol-3-phosphate, pH optima at 6.5, and
KG3Pm in the range of 3-4
mM. The osmotic induction of Gpp2p is blocked in cells that
are defective in the HOG-mitogen-activated protein kinase pathway,
indicating that GPP2 is a target gene for this osmosensing
signal transduction pathway. Together with DOG1 and
DOG2, encoding two highly homologous enzymes that
dephosphorylate 2-deoxyglucose-6-phosphate, GPP1 and
GPP2 constitute a new family of genes for low molecular
weight phosphatases.

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Copyright © 1996 by the American Society for Biochemistry and Molecular Biology.
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